19-nor-testosterone derivatives, their process of preparation and their method of utilization



United States Patent 0 3,136,907 19-NOR-TESTOSTERONE DEREVATZVES, THEE PROCESS OF PREPARATZUN AND THEIR METH- OD 0F UTEAZATION Grard Nomin, Noisy-le-sec, Robert Bucourt, Ciichysous-Buds, and Jean Tessier, Paris, France, assignors to Roussel-UCIAF, S.A., Paris, France, a corporation of France No Drawing. Filed Apr. 10, 1962, Ser. No. 186,341 Claims priority, application France, Apr. 14, 1961, 858,794; Aug. 11, 1961, 870,687

6 Claims. (Cl. 167-74) The present invention relates to the preparation and use of epoxy steroids alkylated in the 2 position. It more particularly relates to Za-lower alkyl-9u,l0a-epoxy-l9- nor-testosterone, ethers and esters thereof, the process of preparing these compounds and the utilization of these compounds.

An object of the present invention is the obtention of 19-nor-testosterone derivatives having the formula:

wherein R represents a lower alkyl; and Z represents a radical selected from the group consisting of hydrogen, lower alkyl and an acyl radical of an organic carboxylic acid having from 2 to 18 carbon atoms.

A further object of the invention is the development of a process for the production of the above 19-nortestosterone derivatives.

Another object of the invention is the obtention of intermediate estradiene derivatives having the formula:

wherein R represents a lower alkyl such as methyl, ethyl,

propyl, isopropyl, etc., Z represents hydrogen; an acyl radical of an organic carboxylic acid having from 2 to 18 carbon atoms; preferably a hydrocarbon carboxylic acid such as alkanoic acids, for example, acetic acid, trimethyl acetic acid; alkenoic acids; cycloalkylalkanoic acids, for example, cyclopentyl acetic acid; benzeneand naphthalene-alkanoic acids, for example, phenyl-acetic acid; benzene carboxylic acids, benzoic acid; cycloalkane carboxylic acids, for example, hexahydrobenzoic acid; etc., or a lower alkyl radical.

The compounds of the invention possess useful phyisological properties. 2u-methyl-9a,l0a-epoxy 19 nor-testosterone possesses particularly an inhibiting action on hypophysis. It can be utilized for the treatment of excesses of hypophysial overstressing of the FSH. factor (folliculostimulating gonadotropic hormone) caused by castration or by menopause. Of course the esters and ethers of 2a-methyl-9a,10wepoxy-l9-nor-testosterone are also useful for the same purpose. I

The process of producing the said 19-nor-testosterone derivatives having the formula:

- wherein R represents a lower alkyl; and Z represents a radical selected from the group consisting of hydrogen, lower alkyl and an acyl radical of an organic carboxylic acid having from 2 to 18 carbon atoms, comprises the steps of subjecting a compound having the formula:

wherein R represents a radical selected from the group consisting of a-lower alkyl and B-lower alkyl and Z has the above-assigned meaning, to the action of a basic cyclization agent in an anhydrous inert organic solvent, separating the estradiene having the formula:

wherein R and Z have the above-assigned meanings, into its 2aand 2li-epimers, reacting the compound having the formula:

wherein R and Z have, the above-assigned meanings with a peracid, and recovering said l9-nor-t'estosterone derivatives.

The corresponding 9a,10a-epoxy-12a-lower alkyl-19 nor-testosterone, or its ester is obtained, depending upon whether the starting product is in the free base form or the esterified form. The final product can be transformed by the usual proceduresinto all other desired functional derivatives.

The starting compounds, the A -3 methyl-7-oxo-8-(3 oxo-2-lower 'alkyl-butyl)-3,4-[3' hydroxy-cyclopentano- (2' ,1) ]-octahydronaphthalene and esters thereof, can be prepared according to the process described in our 00- The 2-lower alkyl-A -estradiene-17B-ol-3-one and es-.

ters thereof are separated into their 20cand 2/3-epimers,

preferably by subjecting them to chromatography over silica gel with elution with methylene chloride containing 0.2% acetone.

The peracid employed in the epoxidation step is preferably an organic percarboxylic acid; for example, peralkanoic acids, such as performic acid, and peracetic acid; and benzene percarboxylic acids, such as perbenzoic acid and perphthalic acid, etc.

The following examples are illustrative of the invention but are not to be deemed limitative. Itisv obvious that other expedients known to the art may be employed. In addition other modes of operation, such as starting with other esters in the 175 position or even the free alcohol or utilizing other 2-lower alkyl derivatives than the Z-methyl deriva te, may be followed.

EXAMPLE I Preparation of the benzoate of 2OL-mthyl-90t,10u-

epoxy-19-n'0r-testoster10ne Step A.--Cycli'zati0 n.5 g. of 176 benzoyloxy 4,5 secO-Z-methyl-A -estrene-3,5-dione (obtained according to United States patent application Serial No. 152,180, now US. Patent No. 3,109,009) were dissolved in 200 cc. of anhydrous toluene. The mixture was cooled to 0 C. and 9.6 cc. of a solution of 1.3 N sodium t-amylate in toluent were added thereto. The mixture was agitated under an atmosphere of nitrogen for a period of 2 hours at 0 C. Thereafter, the reaction mixture was acidified by the addition of dilute acetic acid to a pH of 4. The solution was decanted. The organic phase was washed with water and dried over sodium sulfate. Upon evaporation to dryness under vacuum, a crystalline residue was obtained. This residue consisted of the mixture of 2aand 2,8-epimers of l7fi-benzoyloxy-Z-methyl-A -estradiene-3-one. The product was separated by subjecting it to chromatography over silica gel and elution with methylene culated: C, 76.82%;

chloride containing 0.2% of acetone. This process gave the separation of the two isomers in the 2 position of 17B-benzoyloxy-2-methyl-A -estradiene-3-0ne. 17B-benzoyloxy-h-methyl-A -estradiene-3-one was thus obtained melting at 156.5 C. and having a specific rotation [a] =189 (c.=0.58% in methanol). This compound occurred in the form of cubic crystals, soluble in hot alcohol. I

Analysis.-C H ,O molecular weight=390.5. Calculated: C, 79.96%; H,'7.74%. Found: C, 79.7%; H, 7.7.

U.Vp-spectra (ethanol):

A ,,,.=229230,m EZ: =.484, e =1'8,900 Man ma-303 mu, E;" 3,, =551, e =21,500

On further elution, 17,8-benzoyloxy-2 3-methyl-A estradiene-3-one was obtained melting at C. and having a specific rotation [a] =+1.7 (c.=0,28%' in methanol);

needles soluble in hot alcohol.

U.V. spectra (ethanol):

17,6 benzoyloxy 20c methyl A estradiene 3 one and 17,8-benzoyloxy-Zfl-methyl-A -estradiene-3-one are not described in' the literature.

Sept B.-Ep0xida1i0n.-330 mg. of 17fi-benZoyloxy-2amethyl-A -estradiene-3-one (M.P. 156.5 C.) were introduced into 3.3 cc. of dichlorethane. 0.83 cc. of an ethereal solution containing 30% of perphthalic acid were added and the mixture rformed was allowed to stand 'at room temperature under an atmosphere of nitrogen for a period of 20 hours. The phthalic acid formed was removed by vacuum filtration and then was washed with.

methylene chloride. The organic filtrates were combined, successively washed with an aqueous solution containing 1% of sodium bisulfite, with water, with'a solution containing 5% of sodiumbicar'bonate, and again with water.

,Thereafter the organic solution was dried, filtered and" U.V. spectra (ethanol):

This compound is not described in the literature.

2a-methyl-9u,10a-epoxy-19-nor testosterone, its esters and ethers, can be administered by oral methods, per-t lingual methods, transcutanepps methods or by rectal methods.

The products can be prepared in the form of injectable This compound occurred in the form of- H, 7.44%. Found: C, 77.1%; H,

EXAMPLE II Pharmacological study (a) Determination of the hypophysial inhibitor efiect.- From 30-day old female rats weighing between 75 and 80 g, sisters of the same litters were united 2 by 2 in parabiotic union. One of the rats in the parabiotic union was castrated and treated the same day by the medicine. The animal was treated for a period of days. The animals were sacrificed the eleventh day after the parabiotic union. The genital organs were separated and weighed. In this test the castrated rats received 296 per day of the henzoate of 2u-methyl-9a,1fle-epoxy-19-nor-testosterone for a period of 10 days subcutaneously. The table below gives the results obtained with the difierent lots of rats:

TABLE I Mg. Average weight of the uterus of the entire rat 80 Average weight of the uterus of the castrated rat 88 Average weight of the ovaries of the entire rat 21 Average Weight of the ovaries of the castrated rats (weighed the day of castration, i.e., average weight of the ovaries at the start of the test) 17 A comparison of the average weight of the ovaries of the castrated rats at the start of the experiment taken as the reference weight and the average weight of the ovaries of the entire rat at the end of the experiment shows that there has been no increase in weight of this organ.

In the control couples where the non-treated castrated subject was united parabiotically to the intact subject, the average weight of the ovaries of the intact subject by contrast was equal, at the end of the test, to 150 mg.

It can thus be concluded that the excessive hypophysial overstressing in F.S.H. factor due to the castration is total- .13 inhibited by the benzoate of 2ct-methyl-9oz,l0u-epoxy- 19-nor-testosterone.

(b) Determination of a possible estrogcnic efiect. Histological examination of the vaginas of the castrated females treated with the benzoate of 2a-rnethyl-9a,l0 xepoxy-19-nor testosterone in the previous test of parabiosis showed a vaginal mucification in 75% of the cases. The product possesses thus a weak estrogenic activity.

In contrast, subcutaneous administration to castrated rats weighing about 150 g. for a period of 10 days of a daily dose of 296 of the benzoate of 2a-methyl-9or,1()uepoxy-lQ-nor testosterone caused no caginal mucification.

The dose of 296 per day can thus be considered to constitute the threshold dose for the observation of an estrogenic effect.

In addition, in the Allen and Doisy test, the single administration of 3 mg. of the benzoate of Zea-methyl- 9m,1Oe-epoXy-l9-nor-testosterone was without effect.

(c) Determination of the androgenic and myotrophic efiect.The daily subcutaneous administration of 296 of benzoate of 2a-methyl-9oc,lOar-epuxy-lQ-nor-testosterone for a period of 10 days according to the Herschberger test caused no notable increase of the body weight, of the weight of the seminal vesicules, of weight of the prostate, and of the weight of levator ani, as shown by the table below:

From this test the absence of an androgenic activity and an anabolic activity can be concluded.

(d) Determination of an antilipemic efiect.-Two

TABLE III [Plasmatic turbidity expressed in optical density] Time after ingestion of cream Lot-s 0 3 hours 5 hours 7 hours Control 0. 43 1. 41 0. 75 0. 43 Benzoate oi Qa-methyl-Qa,

sterone 0.71 1. l0 0. 76 0. 52

At the dose of 4 mg. per kg. utilized, the benzoate of 2oz-methyl-9a,l0a-epoxy-l9-nor-testosterone exercised an antilipemic eiiect persisting more than 7 hours after the injection.

The preceding examples are not to be construed as limiting the invention. It is obvious to one skilled in the art that temperatures, the nature of the solvents, the organic carboxylic acid ester or the utilization of other esters, others or the free alcohol, 2oc-methyl-9a,l0oz-epoxyl9-nor-testosterone, may be varied or that equivalent techniques may be employed without departing from the spirit of the invention or the scope of the appended claims.

We claim:

1. The process of preparing 19-nor-testosterone derivatives having the formula:

wherein R represents a lower alkyl; and Z represents a radical selected from the group consisting of hydrogen, lower alkyl and an acyl radical of an organic carboxylic acid having from 2 to-18 carbon atoms, which comprises the steps of subjecting a compound having the formula:

wherein R represents a radical selected from the group consisting of a-lower alkyl and fi-lower alkyl and Z has the above-assigned meaning, to the action of a basic cyclization agent in an anhydrous in'ert organic solvent, sep'a rating-the estradiene having the formulaz-ii I wherein R' and Z have the above-assigned meanings into its 2uand ZB-epimers, reacting the compound having the formula:

wherein R and Z have the above-assigned meanings with a peracid, and recovering said 19-nor-testosterone derivatives. M g I 2. The process of clairn' 1 werein said basicjcycliza tion agent is an alkaline alcoholate. A 7 i 3. The process of claim 2 wherein said alkaline alcoto chromatography to s eparatethe 2aand ZB-Qepimers, reacting the 17p]-benzoyloxy-Za-methyl-A -estradiene-3- one with an organic percarboxylic acid and recovering said benzoate of 2a-methy1-9a,10u-epoxy-19-nor-testosterone.

6; The method of, inhibiting excessive hypophysial overstressing in warm blooded animals which comprises administering a safe but efiective amount of a 19-nor-testosterone derivative having the formula:

wherein R represents a lower alkyl;-and Zrepresents a V radical selected from the group consisting of hydrogen, lower alkyl andan acyl radical of an organic carboxylic acidhaving from 2 to 18 carbon atoms.

References Cited' by the Examiner UNITED STATES PATENTS 3,020,296 2/62 Nomin et a1 260397.45 3,055,885 9/62 Nomin et a1. 260239.5 3,056,811 10/62 Nomin et a1 260-2375 OTHER REFERENCES Aypha'ssorhoet al.: Compt. Rend. Akad. Sci., volume 253, pages 684-681 (July 24, 1961). V

Legrandet al. Co mpt. Rend. Akad. Sci., volume 253, No.21, pages 2378-2380 (November 1, 1961).

Vellu'z et al.: C'ompt. Rend. Akad. Sci., volume 252, N0. 25, 3903-3905 (June 1961).

LEWIS GOTTS, Primary Examiner. 

6. THE METHOD OF INHIBITING EXCESSIVE HYPOPHYSIAL OVERSTRESSING IN WARM BLOODED ANIMALS WHICH COMPRISES ADMINISTERING A SAFE BUT EFFECTIVE AMOUNT OF A 19-NOR-TESTOSTERONE DERIVATIVE HAVING THE FORMULA: 